Neuroprotective activity of thiolutin in epileptic mice: the inhibition of NLRP3 inflammasome.

Neuroinflammation is a key mechanism in epileptogenesis. Thiolutin (THL), an inhibitor of NACHT, LRR, and PYD domains-containing protein 3 (NLRP3) inflammasome, targets a central driver of the neuroinflammatory cascade. However, its role in regulating epileptogenesis and associated pathological changes remains unclear. In this study, we examined the effect of THL on epileptic seizures and neuropathological alterations. Electrode-implanted mice were stimulated with kainic acid (KA) to induce epileptic seizures and subsequently treated with THL, with MCC950 serving as a positive control. The anti-epileptic effects of THL were evaluated by recording brain wave changes and seizure scores, seizure frequency, and total duration within 2 h after KA induction. Cognitive function and neuropathological changes were assessed by the Morris water maze test, novel object recognition test, hematoxylin and eosin staining, Nissl staining, and TUNEL staining. NLRP3 inflammasome activation was analyzed by immunofluorescence and western blot. THL treatment improved brain waves, reduced seizure scores and frequencies, and shortened total seizure duration in KA-induced epileptic mice. THL treatment also reduced neuronal loss, attenuated neuronal apoptosis, and improved cognitive dysfunction in epileptic mice, accompanied by reduced activation of NLRP3 inflammasomes in hippocampal neurons and microglia. These findings suggest that THL exerts neuroprotective effects by blocking NLRP3 inflammasome activation, thereby mitigating neuroinflammatory responses, neuronal injury, and cognitive dysfunction in epileptic mice. Collectively, our results highlight THL as a promising therapeutic agent for epilepsy.